The objective of ABO Grouping
& Rh Typing
To determine ABO grouping and Rh Typing of the given sample by tube method.
Principle
In the ABO Grouping And Rh typing using the antisera is based on the principle of agglutination. Normal human red cells possessing antigens will clump in the presence of the corresponding antibody.
Requirements
1. Blood grouping tubes.
2. Pasteur pipettes
3. Centrifuge
4. Reagents
Anti-A antisera
Anti-B anti-sera
Anti-D antisera
Normal Saline
5. Specimen
EDTA blood
Procedure
1. Prepare 5% Red cell suspension for ABO Grouping and Rh Typing
- 1Mix 5 drops (0.05 ml each) of sediment red
cells with 2 ml normal saline (10 drops of
sediment with 90 drops of normal saline). - Centrifuge at 1500 rpm for 1 to 2 minutes.
Discard the supernatant. Wash 3 times with
normal saline. - Add 4 ml of normal saline to pedimented red
blood cells.
2. Take 3 test tubes. Label them as “A”, “B” and “D”.
3. Place 1 drop of anti-A into the “A” tube, 1 drop of anti-B into the “B” tube, and 1 drop of Anti-D into the “D: tube.
4. Add 1 drop of the RBC suspension to each tube.
5. Gently shake each tube to mix the contents, and then centrifuge tubes at 1500 rpm for 1 minute. The RBCs will form a button or pellet at the bottom of each test tube.
6. Gently resuspend the RBC button and examine for agglutination macro and microscopically.
Observations
Clinical significance
Blood grouping is important in
1. Blood transfusion.
2. To check the identity of birth parents. For example, in a paternity case.
3. Before a person donates blood.
4. Before a person donates an organ for transplantation.
5. Before surgery.
6. To show whether two people could be blood relatives.
7. To check the identity of a person suspected of committing a crime.
Also Read
- Bile Salt (Hay’s Sulphur Method)
- Estimation of occult blood in stool by Benzidine method.
-
“Unlocking the Secrets of Urine: A Comprehensive Guide to Urine Examination”
- Glucosuria (Benedict Method)
- Ketone Body (Rotheras, Gerhardt’s and Strip Method)
- Proteinuria (Heat and acetic acid, Sulphur Salicylic Acid, Hellers or Nitric acid, and Esbach luminometer Method)
- Bence Jones Proteinuria (HCl Method)
- Urobilinogen (Ehrlich Method)
- Bile Pigment (Fouchets Method)
- Occult Blood (Orthotoluidine and Benzidine Method).
- Sedimentation Preparation
- Slide Preparation
- Microscopic Examination.
Reference
1. “Hoffbrand’s Essential Haematology” by A. Victor Hoffbrand, Paul A.H. Moss, and John E. Pettit:
2. “Williams Hematology” by Kenneth Kaushansky, Marshall A. Lichtman, Josef T. Prchal, Marcel M. Levi, Oliver W. Press, and Linda J. Burns
3. “Wint robe’s Clinical Hematology” by John P. Greer, Daniel A. Arber, Bertil E. Glader, Alan F. List, Robert T. Means Jr., Frixos Paraskevas, George M. Rodgers, and Michael H. Schmitt:
“Consultative Hematology: An Issue of Hematology/Oncology Clinics of North America” edited by Mikkael A. Sekeres and Christopher R. Cogle: This book focuses on the consultative aspects of hematology and is a valuable resource for clinicians.